Gladstone Institute of Data Science and Biotechnology, San Francisco, CA, USA.
Graduate Program in Bioengineering, University of California, San Francisco and Berkeley, San Francisco, CA, USA.
Nature. 2022 Aug;608(7921):217-225. doi: 10.1038/s41586-022-04994-6. Epub 2022 Jul 27.
Biological processes depend on the differential expression of genes over time, but methods to make physical recordings of these processes are limited. Here we report a molecular system for making time-ordered recordings of transcriptional events into living genomes. We do this through engineered RNA barcodes, based on prokaryotic retrons, that are reverse transcribed into DNA and integrated into the genome using the CRISPR-Cas system. The unidirectional integration of barcodes by CRISPR integrases enables reconstruction of transcriptional event timing based on a physical record through simple, logical rules rather than relying on pretrained classifiers or post hoc inferential methods. For disambiguation in the field, we will refer to this system as a Retro-Cascorder.
生物过程依赖于基因随时间的差异表达，但对这些过程进行物理记录的方法有限。在这里，我们报告了一个分子系统，用于将转录事件的时间顺序记录到活基因组中。我们通过基于原核反转录酶的工程 RNA 条码来实现这一点，这些条码被反转录成 DNA，并使用 CRISPR-Cas 系统整合到基因组中。CRISPR 整合酶的单向条码整合使得能够根据物理记录通过简单的逻辑规则而不是依赖预先训练的分类器或事后推理方法来重建转录事件的时间。为了在该领域进行区分，我们将这个系统称为 Retro-Cascorder。
